WO2004065398A2 - Synthese et utilisation de nucleosides modifies en n6 et substitues en 2' - Google Patents

Synthese et utilisation de nucleosides modifies en n6 et substitues en 2' Download PDF

Info

Publication number
WO2004065398A2
WO2004065398A2 PCT/US2004/001125 US2004001125W WO2004065398A2 WO 2004065398 A2 WO2004065398 A2 WO 2004065398A2 US 2004001125 W US2004001125 W US 2004001125W WO 2004065398 A2 WO2004065398 A2 WO 2004065398A2
Authority
WO
WIPO (PCT)
Prior art keywords
substituted
group
methyl
alkenyl
nucleophilic
Prior art date
Application number
PCT/US2004/001125
Other languages
English (en)
Other versions
WO2004065398A3 (fr
Inventor
Haoyun An
Kanda Ramasamy
Stephanie Shaw
Original Assignee
Ribapharm Inc.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Ribapharm Inc. filed Critical Ribapharm Inc.
Priority to US10/542,235 priority Critical patent/US7799908B2/en
Publication of WO2004065398A2 publication Critical patent/WO2004065398A2/fr
Publication of WO2004065398A3 publication Critical patent/WO2004065398A3/fr
Priority to US12/876,997 priority patent/US8575331B2/en

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H19/00—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof
    • C07H19/02—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof sharing nitrogen
    • C07H19/04—Heterocyclic radicals containing only nitrogen atoms as ring hetero atom
    • C07H19/16—Purine radicals

Definitions

  • the field of the invention is synthesis of nucleosides, their analogs, and uses thereof.
  • nucleoside analogs wherein such compounds may include modifications on the heterocyclic base as well as on the sugar moiety.
  • modifications are introduced into the already synthesized nucleoside to yield the desired nucleoside analog, while in other approaches the sugar portion is modified and then coupled to the heterocyclic base (which may or may not be modified) to generate the desired nucleoside analog.
  • the desired nucleoside analogs include reactive groups (e.g., OH groups in the sugar, NH 2 group in the heterocyclic base)
  • reactive groups e.g., OH groups in the sugar, NH 2 group in the heterocyclic base
  • modification reagents may react not only with the desired functional group(s) in the heterocyclic base, sugar, and/or nucleoside, but may also modify reactive groups where those are unprotected.
  • relatively high selectivity may be achieved using protecting groups and modification reagents using particular reaction conditions, such conditions may lead to subsequent problems in isolation, isomeric purification, and/or instability of the desired product.
  • the present invention is generally directed to N ⁇ -substituted adenosine nucleoside analogs, and their preparation and use.
  • Especially preferred nucleoside analogs are prepared from a precursor with an electrophilic center that is reacted with a dual nucleophilic reagent under conditions such that the reaction product is formed with high selectivity in which only one of the two nucleophilic groups of the dual nucleophilic reagent reacts with the heterocyclic base.
  • a method of synthesizing an N ⁇ -substituted adenosine analog includes one step in which is provided a dual nucleophilic reagent having a first nucleophilic group and a second nucleophilic group, and an adenosine analog having a leaving group in the 6-position.
  • the dual nucleophilic reagent is reacted with the adenosine under a reaction condition such that the leaving group is replaced by the first nucleophilic group with a selectivity of at least 90%, wherein the reaction condition includes reacting the dual nucleophilic reagent and the heterocyclic base in a non-basic environment under a protective atmosphere and a temperature of at least 40 °C.
  • Further preferred dual nucleophilic reagents will have a structure of R ⁇ R 2 N-NR 3 R 4 - or H 2 N-OR ⁇ , wherein Ri, R 2 , R 3 , and R 4 are independently selected from the group consisting of H, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, heterocycle, substituted heterocycle, acyl, and substituted acyl.
  • Suitable reaction conditions for hydrazine-type reagents will therefore include those in which the non- basic environment comprises at least one of ethanol and methylene chloride, and wherein the temperature is at least 70 °C.
  • suitable reaction conditions for hydroxyamine-type reagents will therefore include those in which the non-basic environment comprises at least one of water and ethanol, and wherein the temperature is at least 70 °C.
  • the step of reacting the dual nucleophilic reagent with the adenosine is performed in a single step and provides a yield of at least 90% of the N6- substituted adenosine analog, and it is still further preferred that the adenosine analog is 6- Chloro-9H-(2'-j8-C-methyl-/3-D-ribofuranosyl)purine, wherein the 2'-/3-C-methyl group of the 6-Chloro-9H-(2'-/3-C-methyl-/3-D-ribofuranosyl)purine is preferably introduced by a step of reacting a protected D-2-ketoribofuranose with methyl magnesium bromide.
  • contemplated compounds obtained by synthetic routes presented herein will include those having a structure according to Formula I:
  • R is selected from the group consisting of NHNH 2; N(CH 3 )1S!H 2 , N(CH 3 )OH, N(CH 2 CH 3 )NH 2 , NHOH, NHOCH 3 , NHOCH 2 CH 3 , NHN(CH 3 ) 2 , N(CH 3 )NHCH 3 , NHNHCH 3 , NHNHOCH 3 , and NHNHCOOCH 3 .
  • Figure 1 is a schematic of one exemplary synthetic route according to the inventive subject matter in which (a) a 2'-modified sugar is prepared using selective deprotection and oxidation of a hydroxy group, (b) the so obtained protected keto-sugar is regioselectively converted to the corresponding beta-substituted sugar, (c) the sugar is coupled to a modified heterocyclic base to form the corresponding modified nucleoside, and (d) the modified nucleoside is reacted with a dual nucleophilic reagent under conditions to provide selective reaction for generation of the desired reaction product.
  • Figure 2 is a schematic of another exemplary synthetic route according to the inventive subject matter in which a protected nucleoside is modified in the 6-position at the heterocyclic base while deprotecting the protected nucleoside with the modification reagent.
  • the inventors discovered a synthetic procedure in which 2'-substituted N 6 -modified nucleosides and their analogs can be prepared in high yields and purity under conditions that favor highly selective reaction of a precursor with only one or tow reactive centers in a dual nucleophilic reagent. Furthermore, contemplated procedures will advantageously reduce undesired oxidation of various desired reaction products.
  • the inventors discovered that suitably protected sugars (e.g. , via benzoyl groups), and particularly furanose sugars can be deprotected at the 2'-position with relatively high selectivity under mild conditions to generate the corresponding 2' -deprotected sugar.
  • the so generated 2'- deprotected sugar is then reacted with a Dess-Martin reagent to selectively oxidize the 2'- hy ⁇ xoxy group to a -keto group ⁇ hile m int inin protection, of the re- inn hydroxy groups in the sugar.
  • the 2'-beta-modified sugar is coupled to a heterocyclic base using DBU and TMSOTf as coupling reagents in large scale to yield the corresponding 2'-modified beta-nucleoside analog in high yield although the 2 '-beta-methyl group generally effects the stereoselectivity for the glycosylation.
  • selective oxidation and alkylation in beta-position using the procedures according to the inventive subject matter results in a substantially stereochemically pure (i.e., at least 90%, more typically at least 95%, and most typically at least 98% ) 2'-beta alkylated product.
  • the so obtained product is then typically re-protected and coupled to a heterocyclic base using standard protocols (see e.g., Protective Groups in Organic Synthesis by Peter G. M. Wuts, Theodora W. Greene, John Wiley & Sons; ISBN: 0471160199).
  • the so produced 2'-beta modified nucleoside analog includes one or more leaving groups at the heterocyclic base that can be modified in a reaction that will retain the remaining reactive groups (protected and/or unprotected) and that will retain the glycosidic bond.
  • An exemplary leaving group, as depicted in Figure 1, is chlorine, which is replaced by a nucleophile in a nucleophilic substitution.
  • heterocycle and “heterocyclic base” are used interchangeably herein and refer to any compound in which a plurality of atoms form a ring via a plurality of covalent bonds, wherein the ring includes at least one atom other than a carbon atom.
  • heterocyclic bases include 5- and 6-membered rings with nitrogen, sulfur, or oxygen as the non-carbon atom (e.g., imidazole, pyrrole, triazole, dihydropyrimidine).
  • heterocylces may be fused (i.e., covalently bound) to another ring or heterocycle, and are thus termed "fused heterocycle" as used herein.
  • fused heterocycles include a 5-membered ring fused to a 6- membered ring (e.g., purine, pyrrolo[2,3-d]pyrimidine), and a 6-membered ring fused to another 6-membered or higher ring (e.g., pyrido[4,5-d]pyrimidine, benzodiazepine).
  • a 6-membered ring e.g., purine, pyrrolo[2,3-d]pyrimidine
  • a 6-membered ring fused to another 6-membered or higher ring e.g., pyrido[4,5-d]pyrimidine, benzodiazepine
  • heterocyclic bases may be aromatic, or may include one or more double or triple bonds.
  • contemplated heterocyclic bases may further include one or more substituents other than hydrogen, and especially contemplated substituents include those referenced below.
  • Contemplated heterocycles or substituted heterocycles are typically attached directly to nucleoside bases or sugars, but coupling of the heterocyclic base to the sugar may also include a linker moiety with at least 1-4 atoms between the heterocyclic base and the sugar.
  • sugar refers to all carbohydrates and derivatives thereof, wherein particularly contemplated derivatives include deletion, substitution or addition of a chemical group in the sugar.
  • particularly contemplated deletions include 3'-deoxy sugars.
  • Especially contemplated substitutions include replacement of the ring-oxygen with sulfur, methylene, or nitrogen, or replacement of a hydroxyl group with a halogen, an amino-, sulfhydryl-, or methyl group, and especially contemplated additions include methylene phosphonate groups.
  • Further contemplated sugars also include sugar analogs (i.e., not naturally occurring sugars), and particularly carbocyclic ring systems.
  • carbocyclic ring system refers to any molecule in which a plurality of carbon atoms form a ring, and in especially contemplated carbocyclic ring systems the ring is formed from 3, 4, 5, or 6 carbon atoms. Examples of these and further preferred sugars are given below.
  • alkyl and “unsubstituted alkyl” are used interchangeably herein and refer to any linear, branched, or cyclic hydrocarbon in which all carbon-carbon bonds are single bonds.
  • substituted alkyl refers to any alkyl that further comprises a functional group, and particularly contemplated functional groups include nucleophilic (e.g., -NH 2 , -OH, -SH- -NC, etc.) and electrophilic groups (e.g., C(O)OR, C(X)OH, etc.), polar groups (e.g., -OH), non-polar groups (e.g., aryl, alkyl, alkenyl, alkynyl, etc.), ionic groups (e.g., -NH 3 + ), and halogens (e.g., -F, -Cl), and all chemically reasonable combinations thereof.
  • nucleophilic e.g., -NH 2 , -OH, -SH- -NC
  • alkenyl and “unsubstituted alkenyl” are used interchangeably herein and refer to any linear, branched, or cyclic alkyl with at least one carbon-carbon double bond.
  • substituted alkenyl refers to any alkenyl that further comprises a functional group, and particularly contemplated functional groups include those discussed above.
  • alkynyl and “unsubstituted alkynyl” are used interchangeably herein and refer to any linear, branched, or cyclic alkyl or alkenyl with at least one carbon-carbon triple bond.
  • substituted alkynyl refers to any alkynyl that further comprises a functional group, and particularly contemplated functional groups include those discussed above.
  • aryl and “unsubstituted aryl” are used interchangeably herein and refer to any aromatic cyclic, alkenyl, or alkynyl.
  • substituted aryl refers to any aryl that further comprises a functional group, and particularly contemplated functional groups include those discussed above.
  • alkaryl is employed where the aryl is further covalently bound to an alkyl, alkenyl, or alkynyl.
  • substituted as used herein also refers to a replacement of a chemical group or substituent (typically H or OH) with a functional group
  • functional groups include nucleophilic (e.g., -NH 2 , -OH, -SH, -NC, etc.) and electrophilic groups (e.g., C(O)OR, C(X)OH, etc.), polar groups (e.g., -OH), non-polar groups (e.g., aryl, alkyl, alkenyl, alkynyl, etc.), ionic groups (e.g., - H 3 + ), and halogens (e.g., -F, -Cl), and all chemically reasonable combinations thereof.
  • nucleophilic e.g., -NH 2 , -OH, -SH, -NC, etc.
  • electrophilic groups e.g., C(O)OR, C(X)OH, etc.
  • polar groups e.g.
  • nucleophilic reagents are considered suitable for use herein.
  • particularly preferred nucleophilic reagents include compounds in which two or more nucleophilic centers are present (i.e., dual nucleophilic reagents).
  • dual nucleophilic reagents include those in which the first nucleophilic group is a primary or secondary amine group, and in which the second nucleophilic group is a hydroxyl group, ether group, or a primary or secondary amine.
  • nucleophilic reagents include various substituted and unsubstituted hydrazines and hydroxyamines with the general formula R ⁇ R 2 N-NR 3 R and R ⁇ R 2 N-OR 5 , in which Ri, R 2 , R 3 , R 4 , and R 5 are independently selected from the group consisting of H, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, heterocycle, substituted heterocycle, acyl, and substituted acyl.
  • Exemplary dual nucleophilic reagents therefore include NH 2 NH 2 , H 2 N(CH 3 ) 2 , NHCH 3 NHCH 3 , NH 2 NHCH 3 , NH 2 NHC(O)CH 3 , NH 2 NHC(O)OCH 3 , NH 2 OCH 2 CH 3 , NH 2 NHR, NH(Boc)NHR, NHRNHR, NH 2 OH, NHROH, NHROR, NH 2 OR, and NR 2 OH, where R is independently H, C1-C5 alkyl/alkenyl, alkenyl, or C5-C15 aryl.
  • contemplated synthetic protocols will provide substantially stereoselective pure (i.e., at least 90%, more typically at least 95%, and most typically at least 98%) reaction products.
  • the dual nucleophilic reagent is a monosubstituted hydrazine of the general formula R ⁇ HN-NH 2 (with Ri as defined above)
  • the nitrogen of the substituted amino group will act as the nucleophilic group under conditions according to the inventive subject matter.
  • a person of ordinary skill in the art would rather expect that the nitrogen atom of the NH2 group would act (at least to some extent) as a nucleophilic center due to reduced steric hindrance.
  • this is clearly not the case.
  • the dual nucleophilic reagent is a monosubstituted hydroxyamine of the general formula R ⁇ O-NH 2 , HO-NRiH, or R ⁇ O-NR 2 H (with Ri and R 2 as defined above)
  • the nitrogen of the amino group will act as the nucleophilic group under conditions according to the inventive subject matter, while the oxygen will typically not react to a significant degree (i.e., less than 5%, more typically less than 3%, and most typically less than 2%).
  • the inventors discovered that in reaction protocols according to the inventive' subject matter the OH group is more nucleophilic than an NH 2 group under strong basic conditions, but that the NH 2 group is more nucleophilic than OH under weak basic conditions, and the inventors eventually discovered that the nitrogen (substituted where substituted hydrazine is used, or nitrogen in hydroxyamines) reacted first and exclusively with the nucleoside electrophile without adding any base.
  • the inventors further observed that under such reaction conditions the reaction conditions for the dual nucleophilic reagent also provide a deprotection of the protecting groups at the sugar (which can take place in the same reaction mixture in the same vessel).
  • the nucleoside analog after coupling the sugar to the heterocyclic base, is first deprotected and that the deprotected nucleoside analog is then reacted with the nucleophilic reagent to yield the desired deprotected 2'- methyl-6-substituted adenosine.
  • the nucleophilic reagent may also be employed to remove one or more of the protecting groups at the same time in the step of replacing the leaving group as depicted in Figure 2.
  • the 6-position modifying agent i.e., the nucleophilic agent
  • This second strategy saves one step, and gives a high quality, high yield product that is easily purified. Therefore, this later strategy is superior to the first strategy, although both approaches are effectively utilized for the synthesis of this type of derivatives.
  • contemplated synthetic routes provide selective and mild reactive conditions to form the 2'-modified sugar, while at the same time significantly reducing oxidative damage to the compounds by employing the 6- position modifying agent as a deprotectant when compared to other methods. Consequently, the inventors contemplate a method of synthesizing an N6-substituted adenosine analog in which in one step a dual nucleophilic reagent is provided having a first nucleophilic group and a second nucleophilic group, and an adenosine analog having a leaving group in 6-position.
  • the dual nucleophilic reagent is reacted with the adenosine under a reaction condition such that the leaving group is replaced by the first nucleophilic group with a selectivity of at least 90% (more preferably at least 95%, even more preferably at least 97%) and wherein the reaction condition comprises a reaction of the dual nucleophilic reagent with the adenosine in a non-basic environment under a protective atmosphere and a temperature of at least 40 °C.
  • adenosine analog generally refers to a nucleoside in which the nucleoside has a sugar that is coupled to a heterocyclic base.
  • Preferred heterocyclic bases in contemplated adenosine analogs will have a purine scaffold, in which one or more nitrogen atoms are optionally replaced by a (substituted) carbon atom, and in which the sugar is substituted at the 2'-position (preferably in beta orientation).
  • a N ⁇ -substituted adenosine analog is a purine nucleoside in which the purine moiety has a substituted amino group in 6-position of the purine scaffold.
  • non-basic environment refers to an environment for reacting the heterocyclic base with a first nucleophilic group of the dual nucleophilic reagent under conditions that either entirely exclude addition of a base, or maintain addition of a base to the reaction medium below a level that would enable the undesirable reaction of the second nucleophilic group to form at least 10% (and more typically at least 5%) product in which the second group has reacted with the heterocyclic base.
  • protective atmosphere refers to any atmosphere that replaces ambient air in the reaction vessel with a gas substantially depleted (i.e., less than 1 vol%, more typically less than 0.1 vol%) of oxygen.
  • suitable protective atmospheres include pure argon, nitrogen, or helium.
  • the dual nucleophilic reagent has a structure of R ⁇ N- R ⁇ , with Ri, R 2 , R 3 , and R* independently selected from the group consisting of H, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, heterocycle, substituted heterocycle, acyl, and substituted acyl
  • the non-basic environment comprises at least one of ethanol and methylene chloride, and that the temperature is at least 70 °C.
  • the dual nucleophilic reagent has a structure of H 2 N-OR ⁇ , and wherein Ri is selected from the group consisting of H, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, heterocycle, substituted heterocycle, acyl, and substituted acyl
  • Ri is selected from the group consisting of H, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, heterocycle, substituted heterocycle, acyl, and substituted acyl
  • the non-basic environment comprises at least one of water and ethanol, and wherein the temperature is at least 70 °C.
  • R is selected from the group consisting of NHNH j N(CH 3 )NH 2 , N(CH 3 )OH, N(CH 2 CH 3 )NH 2 , NHOCH 3 , NHN(CH 3 ) 2 , N(CH 3 )NHCH 3 , NHNHCH 3 , NHNHCOOCH 3 , NHNHOCH 3 , NHOCH 2 CH 3 , and NHOH.
  • especially preferred compounds include 2'- ⁇ -C-Methyl-N ⁇ -amino-N ⁇ -methyl- ⁇ -D-ribo ranosyladenosine, N ⁇ -Ethyl-N 5 -amino-9H- (2'- ⁇ -C-methyl- ⁇ -D-ribo-manosyl)purine, N tf - ⁇ ydroxy-N 5 -methyl-9H-(2'- ⁇ -C-methyl- y 5-D- ribofuranosyl)purine, N tf - ⁇ ydroxy-9H-(2'- ⁇ -C-methyl- ⁇ -D-ribofuranosyl)purine, and N 6 - Methoxy-9H-(2'- ⁇ -C-methyl--?-D-ribofuranosyl)purine.
  • contemplated synthetic procedures will provide 2- beta substituted- ⁇ 6 -substituted nucleosides and nucleotides that may exhibit numerous biological activities.
  • Especially contemplated biological activities include in vitro and in vivo inhibition of DNA and/or RNA polymerases, reverse transcriptases, and ligases. Therefore, contemplated nucleosides will exhibit particular usefulness as in vitro and/or in vivo antiviral agents, antineoplastic agents and immunomodulatory agents.
  • Particularly contemplated antiviral activities include at least partial reduction of viral titers of respiratory syncytial virus (RSV), hepatitis B virus (HBV), hepatitis C virus (HCV), herpes simplex type 1 and 2, herpes genitalis, herpes keratitis, herpes encephalitis, herpes zoster, human immunodeficiency virus (HIN), influenza A virus, Hanta virus (hemorrhagic fever), human papilloma virus (HPN), yellow fever virus, and measles virus.
  • RSV respiratory syncytial virus
  • HBV hepatitis B virus
  • HCV hepatitis C virus
  • herpes simplex type 1 and 2 herpes simplex type 1 and 2
  • herpes genitalis herpes keratitis
  • herpes encephalitis herpes zoster
  • HIN human immunodeficiency virus
  • influenza A virus Hanta virus (
  • the anti-HCV activity of the nucleosides and libraries can readily be tested by Replicon and BVDV cell-line based assays well known in the art (see e.g., V. Lohmann, F. Korner, J.-O. Koch, U. Herian, L. Theilmann, R. Bartenschlager, "Replication of a Subgenomic Hepatitis C virus R ⁇ As in a Hepatoma Cell Line", Sciences, 1999, 285, 110).
  • Exemplary biological activity data and experimental conditions are described in our copending International patent application with the serial number PCT/US02/34026, which was filed 10/23/02, and which is incorporated by reference herein.
  • Especially contemplated immunomodulatory activity includes at least partial reduction of clinical symptoms and signs in arthritis, psoriasis, inflammatory bowel disease, juvenile diabetes, lupus, multiple sclerosis, gout and gouty arthritis, rheumatoid arthritis, rejection of transplantation, giant cell arteritis, allergy and asthma, but also modulation of some portion of a mammal's immune system, and especially modulation of cytokine profiles of Type 1 and Type 2.
  • modulation of Type 1 and Type 2 cytokines may include suppression of both Type 1 and Type 2, suppression of Type 1 and stimulation of Type 2, or suppression of Type 2 and stimulation of Type 1.
  • nucleosides are administered in a pharmacological composition
  • suitable nucleosides can be formulated in admixture with a pharmaceutically acceptable carrier.
  • contemplated nucleosides can be administered orally as pharmacologically acceptable salts, or intravenously in a physiological saline solution (e.g., buffered to a pH of about 7.2 to 7.5).
  • physiological saline solution e.g., buffered to a pH of about 7.2 to 7.5.
  • Conventional buffers such as phosphates, bicarbonates or citrates may be used for this purpose.
  • one of ordinary skill in the art may modify the formulations within the teachings of the specification to provide numerous formulations for a particular route of administration.
  • contemplated nucleosides may be modified to render them more soluble in water or other vehicle, which for example, may be easily accomplished with minor modifications (salt formulation, esterification, etc.) that are well within the ordinary skill in the art. It is also well within the ordinary skill of the art to modify the route of administration and dosage regimen of a particular compound in order to manage the pharmacokinetics of the present compounds for maximum beneficial effect in a patient.
  • prodrug forms of contemplated nucleosides maybe formed for various purposes, including reduction of toxicity, increasing the organ or target cell specificity, etc.
  • acylated (acetylated or other) derivatives, pyridine esters and various salt forms of the present compounds are preferred.
  • One of ordinary skill in the art will recognize how to readily modify the present compounds to pro-drug forms to facilitate delivery of active compounds to a target site within the host organism or patient.
  • One of ordinary skill in the art will also take advantage of favorable pharmacokinetic parameters of the pro-drug forms, where applicable, in delivering the present compounds to a targeted site within the host organism or patient to maximize the intended effect of the compound.
  • contemplated compounds may be administered alone or in combination with other agents for the treatment of various diseases or conditions.
  • Combination therapies according to the present invention comprise the administration of at least one compound of the present invention or a functional derivative thereof and at least one other pharmaceutically active ingredient.
  • the active ingredient(s) and pharmaceutically active agents may be administered separately or together and when administered separately this may occur simultaneously or separately in any order.
  • the amounts of the active ingredient(s) and pharmaceutically active agent(s) and the relative timings of administration will be selected in order to achieve the desired combined therapeutic effect.
  • the pot temperature of the thick orangish-amber suspension was increased to 70 °C, and the condition was maintained for a 3.5h time period.
  • the reaction mixture was allowed to cool to ambient conditions.
  • the reaction mixture was cooled to — 1°C and maintained for 0.5 h.
  • the filter cake was washed with 1L of water, followed by 2 x 100 ml of ethyl alcohol, and then finally with 2 x 150 ml of diethyl ether.
  • the cake was dried at high vacuum ( ⁇ 0. lmmHg)/ambient temperature for 24 h. 101.56 g of white powder was obtained in 90% yield.
  • a 1L 3 neck RB flask was fitted with a mechanical stirrer, a heating mantle, a thermometer with adapter, and a nitrogen inlet/outlet.
  • the vessel was charged with 400 ml (4.24 mol) of acetic anhydride (4ml/g) and 0.5 g (0.00263 mol) of p-toluenesulfonic acid- monohydrate under nitrogen atmosphere and stirring.
  • the reaction mixture was charged with 100 g (0.357 mol) of l-hydroxy-l,2-benziodoxol-3-(lH)-one.
  • the reaction mixture was allowed to cool to ambient temperature and then cooled to -2 °C.
  • the white suspension was stirred at -2 °C for 0.5 h and filtered.
  • the filter cake was washed with 5 x 50 ml of diethyl ether and then quickly transferred to an amber bottle under an argon atmosphere. The bottle was subsequently stored under refrigeration at ⁇ 5°C. 136.27 g of the desired product was obtained as a white solid in 90% yield.
  • step 1 Synthesis of l',3',5'-Tri-0-benzoyl-D-ribofuranose (2) (step 1): Compound 2 was synthesized by a modified procedure based on the literature [Brodfuehrer, P. R.; Sapino, C, Jr.; Howell, H. G. J. Org. Chem. 1985, 50, 2598].
  • step 2 Synthesis of l ⁇ 3',5'-Tri-O-benzoyl-D-2-Ketoribofuranose 4 (step 2): A 3L 3- necked RB flask was fitted with a mechanical stirrer, a thermometer with adapter, and a nitrogen inlet/outlet, to which was added 201 g (0.474 mol) of 1,1,1 -triacetoxy- 1,1 -dihydro- l,2-benziodoxol-3-(lH)-one (Dess-Martin reagent, 3) and 1000 mL (15.60 mol) of dichloromethane under nitrogen atmosphere.
  • step 2 A 3L 3- necked RB flask was fitted with a mechanical stirrer, a thermometer with adapter, and a nitrogen inlet/outlet, to which was added 201 g (0.474 mol) of 1,1,1 -triacetoxy- 1,1 -dihydro- l,2-benziodoxol-3-(
  • reaction mixture was cooled to -1 °C, and 100 g (0.216 mol) of l',3',5'-tri-O-ber ⁇ zoyl-D-ribofuranose (2) were added.
  • the resulting reaction mixture was stirred at room temperature for 24 h and concentrated.
  • the resultant residue was triturated with diethyl ether.
  • the resulting ether triturate was filtered through a pad of Celite, and then treated with 1L of 1.0 M sodium thiosulfate solution.
  • the organic phase was washed with sodium thiosulfate solution, and saturated sodium bicarbonate solution followed by brine.
  • the organic phase was dried over magnesium sulfate and concentrated.
  • the clear viscous pale yellow oily residue was subsequently dissolved in 2 L of dichloromethane.
  • the solution was further treated with 500g of magnesium sulfate for 24 h and concentrated.
  • the residue was further dried under high vacuum to provide 95.47 g (96%) of the desired product 4 as a white foam. [Cook, G. P.; Greenberg, M. M. J. Org. Chem. 1994, 59, 4704-4706].
  • the reaction mixture was allowed to slowly warm to a pot temperature of -30 °C at which point 100 g (0.217 mol) of l,3,5-tri-O-benzoyl-alpha-D-2-keto-ribofuranose in 200 ml of diethyl ether (2ml/g) was added drop wise.
  • the reaction mixture was allowed to stir at -30 °C for a 4 h.
  • the organic phase was separated, and the aqueous phase was extracted with 3 x 2000 ml of diethyl ether.
  • the combined organic phase was washed with water and then dried over magnesium sulfate.
  • a 2L 3 neck RB flask was fitted with a mechanical stirrer, a thermometer with adapter, an addition funnel, and a nitrogen inlet/outlet to which were added 6.63 g (0.0543 mol) of 4-dimethlyaminopyridine and 500 ml (7.80 mol) of dichloromethane under nitrogen atmosphere. 85 ml (0.613 mol) of triethylamine were added followed by the addition of 12.6 ml (0.1086 mol) of benzoyl chloride drop wise. 25.87 g (0.0543 mol) of sugar intermediates 5 and 6 obtained above in 125ml of dichloromethane (5ml/g) were added drop wise.
  • the resulting clear light amber reaction mixture was allowed to stir at ambient conditions for 3h to complete the reaction (TLC analysis on silica gel, 4:1 Hex/EtOAc).
  • the reaction mixture was diluted with 2.5L of diethyl ether, and the clear pale amber solution was partitioned with a 750ml portion of 1 Molar HC1 solution in an extraction vessel.
  • the organic phase was separated and washed with 2 x 500 ml of 1 Molar HC1 solution, followed by a 500 ml of water and 2 x 500ml portions of saturated sodium bicarbonate solution.
  • the organic solution was dried over sodium sulfate and concentrated. The remaining residue was subsequently pumped at high vacuum/ambient temperature for a 14 h.
  • the reaction mixture was then warmed to room temperature during a 30 minute period.
  • the resulting reaction mixture was heated at 60 °C for 4 h and concentrated to dryness.
  • the residue was partitioned between ethyl acetate and saturated NaHCO 3 ( 300/200 ml).
  • the organic phase was separated, and the aqueous phase was extracted in ethyl acetate.
  • the combined organic phase was washed with brine, dried over anhydrous sodium sulfate, and concentrated to dryness.
  • the resultant residue was purified by flash chromatography on a silica gel column using hexane -» EtO Ac as the eluent. The pure fractions were collected and concentrated to dryness to provide 6.60 g (95%) of the titled compound 8.
  • 6-Chloro-9H-(2'- ⁇ -C-methyl->S-D-ribofuranosyl)purine (9) (step 6): 6-Chloro-9H- (2'- ⁇ -C-methyl-2',3 5'-tri-O-benzoyl- ⁇ -D-ribofuranosyl) ⁇ urine (8) (6.5 g, 10.4 mmol) was dissolved in C ⁇ C1 3 (50 ml) and placed in a steel bomb. To this solution was added MeOH / NH (200 ml). The bomb was closed and allowed to stir at room temperature for 8 h. The bomb was cooled to 0 °C and opened carefully. The reaction mixture was concentrated to dryness.
  • step 7 A hot solution of 6-chloro-9H-(2'- ⁇ -C-methyl-yf?-D-ribofuranosyl)purine (9) (1.2 g, 4.0 mmol) in EtO ⁇ (100 ml) was diluted with C ⁇ C1 3 (40 ml). The resulting solution was cooled to room temperature. To this stirred solution was added N-methylhydrazine (1.06 ml, 20 mmol) under an Argon atmosphere. The reaction mixture was allowed to stir at room temperature for 12 h and concentrated to dryness.
  • I'- ⁇ -C-Methyl-iV ⁇ -amitto-iV ⁇ -methyl-jS-D-ribofuranosyladenosine (10) (step 8): A mixture of 6-chloro-9H-(2'- ⁇ -C-methyl-2',3',5'-tri-O-benzoyl- y r-?-D-ribofuranosyl)purine (8) (270 mg, 0.43 mmol) and N-methylhydrazine (10 mL) in EtO ⁇ (8 mL) was stirred at 90 °C (bath temperature) under argon atmosphere for 12 h. The reaction mixture was cooled and concentrated to dryness.
  • the resultant residue was purified by flash chromatography on a silica gel column using a gradient of C ⁇ Cl -MeO ⁇ (20:1 to 2:1) as eluent to give 122 mg (92%) of pure product 10 as a white foam.
  • the product obtained this way shows the same spectroscopic properties as that obtained by the alternative procedure shown above.
  • N 5 -Ethyl-iV ⁇ f -amino-9H-(2'- ⁇ -C-methyl-y0-D-ribofuranosyl)purine (11) (step 9): A mixture of 6-chloro-9H-(2'- ⁇ -C-methyl-2' ,3 ' ,5 '-tri-O-benzoyl-/?-D-ribofuranosyl)purine (8) (0.43 mmol) and N-ethylhydrazine (10 mL) in EtO ⁇ (8 L) was stirred at 90 °C (bath temperature) under argon atmosphere for 12 h. The reaction mixture was cooled and concentrated to dryness.
  • 3-D-ribofuranosyl) purine (11) can be prepared by the following procedure: Ethylhydrazine oxalate (4.15 g, 27.7 mmol) was suspended in EtOH (50 ml) and treated with N,N-diisopropylethylamine (9.56 ml, 55.00 mmol).
  • ⁇ -Hydroxy-N 5 -methyl-9H-(2'- ⁇ -C-methyl- ⁇ D-ribofuranosyl)purine (12) (step 10): A mixture of 6-chloro-9H-(2'- ⁇ -C-methyl-2',3',5'-tri-O-benzoyl-)3-D- ribofuranosyl)purine (8) (0.43 mmol) and N-methylhydroxyamine (2.5 mmol) in EtO ⁇ (20 L) was stirred at 90 °C bath temperature under argon atmosphere for 12 h. The reaction mixture was cooled and concentrated to dryness.
  • N f -Methoxy-9H-(2'- ⁇ -C-methyl- ⁇ -D-ribofuranosyl)purine (14) (step 12).
  • a mixture of 6-chloro-9H-(2'- ⁇ -C-methyl-2',3 ',5 '-tri-O-benzoyl-yt-?-D-ribofuranosyl)purine (8) (0.43 mmol) and O-methylhydroxyamine (2.5 mmol) in EtO ⁇ (20 mL) was stirred at 90 °C (bath temperature) under argon atmosphere for 12 h. The reaction mixture was cooled and concentrated to dryness.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Biochemistry (AREA)
  • Biotechnology (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Molecular Biology (AREA)
  • Saccharide Compounds (AREA)

Abstract

L'invention concerne un procédé perfectionné pour préparer un analogue de nucléoside à sucre modifié. Ce procédé comporte un protocole dans lequel un groupe hydroxy d'un sucre est sélectivement libéré de sa protection et oxydé avant la modification nucléophilique du groupe carbonyle correspondant. Le sucre modifié est alors couplé à une base hétérocyclique, qui est modifiée par un réactif nucléophile lors d'une opération ultérieure, laquelle permet d'obtenir des analogues d'adénosine modifiés en N6 à stéréosélectivité élevée.
PCT/US2004/001125 2003-01-15 2004-01-15 Synthese et utilisation de nucleosides modifies en n6 et substitues en 2' WO2004065398A2 (fr)

Priority Applications (2)

Application Number Priority Date Filing Date Title
US10/542,235 US7799908B2 (en) 2003-01-15 2004-01-15 Synthesis and use of 2′-substituted-N6 -modified nucleosides
US12/876,997 US8575331B2 (en) 2003-01-15 2010-09-07 Synthesis and use of 2′-substituted-N6-modified nucleosides

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US44066603P 2003-01-15 2003-01-15
US60/440,666 2003-01-15

Related Child Applications (2)

Application Number Title Priority Date Filing Date
US10/542,235 A-371-Of-International US7799908B2 (en) 2003-01-15 2004-01-15 Synthesis and use of 2′-substituted-N6 -modified nucleosides
US12/876,997 Continuation US8575331B2 (en) 2003-01-15 2010-09-07 Synthesis and use of 2′-substituted-N6-modified nucleosides

Publications (2)

Publication Number Publication Date
WO2004065398A2 true WO2004065398A2 (fr) 2004-08-05
WO2004065398A3 WO2004065398A3 (fr) 2005-03-03

Family

ID=32771846

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2004/001125 WO2004065398A2 (fr) 2003-01-15 2004-01-15 Synthese et utilisation de nucleosides modifies en n6 et substitues en 2'

Country Status (2)

Country Link
US (2) US7799908B2 (fr)
WO (1) WO2004065398A2 (fr)

Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2004065398A3 (fr) * 2003-01-15 2005-03-03 Ribapharm Inc Synthese et utilisation de nucleosides modifies en n6 et substitues en 2'
US7094768B2 (en) 2002-09-30 2006-08-22 Genelabs Technologies, Inc. Nucleoside derivatives for treating hepatitis C virus infection
US7323449B2 (en) 2002-07-24 2008-01-29 Merck & Co., Inc. Thionucleoside derivatives as inhibitors of RNA-dependent RNA viral polymerase
US7425547B2 (en) 2002-09-30 2008-09-16 Genelabs Technologies, Inc. Nucleoside derivatives for treating hepatitis C virus infection
JP2015535261A (ja) * 2012-10-29 2015-12-10 コクリスタル ファーマ，インコーポレイテッド ウイルス感染及び癌の治療のためのピリミジンヌクレオチド及びその一リン酸プロドラッグ
US9968628B2 (en) 2000-05-26 2018-05-15 Idenix Pharmaceuticals Llc Methods and compositions for treating flaviviruses and pestiviruses
US10363265B2 (en) 2000-05-23 2019-07-30 Idenix Pharmaceuticals Llc Methods and compositions for treating hepatitis C virus
US10525072B2 (en) 2002-11-15 2020-01-07 Idenix Pharmaceuticals Llc 2′-branched nucleosides and flaviviridae mutation

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7217815B2 (en) * 2002-01-17 2007-05-15 Valeant Pharmaceuticals North America 2-beta -modified-6-substituted adenosine analogs and their use as antiviral agents

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2003062256A1 (fr) * 2002-01-17 2003-07-31 Ribapharm Inc. Analogues d'adenosine 2'-beta-modifiee-6-substituee et leur utilisation en tant qu'agents antiviraux

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
NZ536123A (en) * 2002-05-06 2006-09-29 Genelabs Tech Inc Nucleoside derivatives for treating hepatitis C virus infection
FR2845034B1 (fr) * 2002-09-27 2006-01-13 Valeo Climatisation Installation de climatisation munie d'un dispositif de detection et d'estimation de l'humidite de l'air
AU2003279797B2 (en) 2002-09-30 2009-10-22 Genelabs Technologies, Inc. Nucleoside derivatives for treating hepatitis C virus infection
US7799908B2 (en) * 2003-01-15 2010-09-21 Valeant Pharmaceuticals North America Synthesis and use of 2′-substituted-N6 -modified nucleosides

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2003062256A1 (fr) * 2002-01-17 2003-07-31 Ribapharm Inc. Analogues d'adenosine 2'-beta-modifiee-6-substituee et leur utilisation en tant qu'agents antiviraux

Cited By (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US10363265B2 (en) 2000-05-23 2019-07-30 Idenix Pharmaceuticals Llc Methods and compositions for treating hepatitis C virus
US10758557B2 (en) 2000-05-23 2020-09-01 Idenix Pharmaceuticals Llc Methods and compositions for treating hepatitis C virus
US9968628B2 (en) 2000-05-26 2018-05-15 Idenix Pharmaceuticals Llc Methods and compositions for treating flaviviruses and pestiviruses
US7323449B2 (en) 2002-07-24 2008-01-29 Merck & Co., Inc. Thionucleoside derivatives as inhibitors of RNA-dependent RNA viral polymerase
US7094768B2 (en) 2002-09-30 2006-08-22 Genelabs Technologies, Inc. Nucleoside derivatives for treating hepatitis C virus infection
US7425547B2 (en) 2002-09-30 2008-09-16 Genelabs Technologies, Inc. Nucleoside derivatives for treating hepatitis C virus infection
US7432248B2 (en) 2002-09-30 2008-10-07 Genelabs Technologies, Inc. Nucleoside derivatives for treating Hepatitis C virus infection
US7629328B2 (en) 2002-09-30 2009-12-08 Smithkline Beecham Corporation Nucleoside derivatives for treating hepatitis C virus infection
US7629320B2 (en) 2002-09-30 2009-12-08 Smithkline Beecham Corporation Nucleoside derivatives for treating hepatitis C virus infection
US10525072B2 (en) 2002-11-15 2020-01-07 Idenix Pharmaceuticals Llc 2′-branched nucleosides and flaviviridae mutation
WO2004065398A3 (fr) * 2003-01-15 2005-03-03 Ribapharm Inc Synthese et utilisation de nucleosides modifies en n6 et substitues en 2'
JP2015535261A (ja) * 2012-10-29 2015-12-10 コクリスタル ファーマ，インコーポレイテッド ウイルス感染及び癌の治療のためのピリミジンヌクレオチド及びその一リン酸プロドラッグ

Also Published As

Publication number Publication date
US20060135465A1 (en) 2006-06-22
US7799908B2 (en) 2010-09-21
US20110196144A1 (en) 2011-08-11
WO2004065398A3 (fr) 2005-03-03
US8575331B2 (en) 2013-11-05

Similar Documents

Publication Publication Date Title
US8575331B2 (en) Synthesis and use of 2′-substituted-N6-modified nucleosides
EP0646125B1 (fr) Analogues de nucleosides a base de 1,5-anhydrohexitol et leur utilisation pharmaceutique
US20040266996A1 (en) Methods of manufacture of 2'-deoxy-beta-L-nucleosides
JPH0637394B2 (ja) 腫瘍治療剤
JP2003176296A (ja) 単環式ヌクレオシド、その類似体および使用
EP1804812A2 (fr) Nucleosides de pyrroloý2,3-d¨pyrimidine fluores destines au traitement d'infections ribovirales dependantes de l'arn
CA2706432A1 (fr) Composes nucleosidiques antiviraux
WO1988007532A1 (fr) Derives d'oxyde de 2',3' didesoxyribofuranne
WO2003062255A2 (fr) Nucleosides a sucre modifie utilises en tant qu'inhibiteurs de la replication virale
WO2003062256A1 (fr) Analogues d'adenosine 2'-beta-modifiee-6-substituee et leur utilisation en tant qu'agents antiviraux
JPH05213941A (ja) 双環式ヌクレオシド、オリゴヌクレオチド、それらの製造方法及び中間体類
ZA200505040B (en) Process for the production of 3'-nucleoside prodrus
CN105979951A (zh) 作为流感rna复制的抑制剂的4’-二氟甲基取代的核苷衍生物
WO2003061385A1 (fr) Composes et banques de nucleosides tricycliques, synthese et utilisation comme agents antiviraux
WO1997037993A1 (fr) DERIVES DE 9-(2-DESOXY-2-FLUORO-4-THIO-β-D-ARABINOFURANOSYL)PURINE
JP2017057200A (ja) 抗ｄｎａウィルス活性などの生理活性を有するヌクレオシド誘導体
KR20070073958A (ko) 디플루오로뉴클레오시드 및 이의 제조 방법
WO1999043690A1 (fr) Compose de l-4'-arabinofuranonucleoside et composition medicinale le contenant
US20040039190A1 (en) Process for the preparation of 9-beta-anomeric nucleoside analogs
JPH09249690A (ja) ４’−チオアラビノプリンヌクレオシド
EP1253154B1 (fr) Purification de nucléosides de 2'-deoxypurine protégés en 5'
Schirmeister‐Tichy et al. Nucleotides, Part LIX, Synthesis, Characterization, and Biological Activities of New Potential Antiviral Agents:(2′–5′) Adenylate Trimer Analogs Containing 3′‐Deoxy‐3′‐(hexadecanoylamino) adenosine at the 2′‐Terminus
WO2006122207A1 (fr) 6-hydrazinopurine 2'-methyl ribonucleosides et nucleotides pour le traitement de hcv
US5283331A (en) 2-halogeno-oxetanocin A and phosphoric ester thereof
JP6983814B2 (ja) 抗ウイルス活性を示すヌクレオシド誘導体

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A2

Designated state(s): AE AG AL AM AT AU AZ BA BB BG BR BW BY BZ CA CH CN CO CR CU CZ DE DK DM DZ EC EE EG ES FI GB GD GE GH GM HR HU ID IL IN IS JP KE KG KP KR KZ LC LK LR LS LT LU LV MA MD MG MK MN MW MX MZ NA NI NO NZ OM PG PH PL PT RO RU SC SD SE SG SK SL SY TJ TM TN TR TT TZ UA UG US UZ VC VN YU ZA ZM ZW

AL Designated countries for regional patents

Kind code of ref document: A2

Designated state(s): BW GH GM KE LS MW MZ SD SL SZ TZ UG ZM ZW AM AZ BY KG KZ MD RU TJ TM AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HU IE IT LU MC NL PT RO SE SI SK TR BF BJ CF CG CI CM GA GN GQ GW ML MR NE SN TD TG

121 Ep: the epo has been informed by wipo that ep was designated in this application
ENP Entry into the national phase

Ref document number: 2006135465

Country of ref document: US

Kind code of ref document: A1

WWE Wipo information: entry into national phase

Ref document number: 10542235

Country of ref document: US

122 Ep: pct application non-entry in european phase
WWP Wipo information: published in national office

Ref document number: 10542235

Country of ref document: US